Yuan Hua, Perry Cynthia N, Huang Chengqun, Iwai-Kanai Eri, Carreira Raquel S, Glembotski Christopher C, Gottlieb Roberta A
Bioscience Center, San Diego State University, San Diego, CA, USA.
Am J Physiol Heart Circ Physiol. 2009 Feb;296(2):H470-9. doi: 10.1152/ajpheart.01051.2008. Epub 2008 Dec 19.
Bacterial endotoxin lipopolysaccharide (LPS) is responsible for the multiorgan dysfunction that characterizes septic shock and is causal in the myocardial depression that is a common feature of endotoxemia in patients. In this setting the myocardial dysfunction appears to be due, in part, to the production of proinflammatory cytokines. A line of evidence also indicates that LPS stimulates autophagy in cardiomyocytes. However, the signal transduction pathway leading to autophagy and its role in the heart are incompletely characterized. In this work, we wished to determine the effect of LPS on autophagy and the physiological significance of the autophagic response. Autophagy was monitored morphologically and biochemically in HL-1 cardiomyocytes, neonatal rat cardiomyocytes, and transgenic mouse hearts after the administration of bacterial LPS or TNF-alpha. We observed that autophagy was increased after exposure to LPS or TNF-alpha, which is induced by LPS. The inhibition of TNF-alpha production by AG126 significantly reduced the accumulation of autophagosomes both in cell culture and in vivo. The inhibition of p38 MAPK or nitric oxide synthase by pharmacological inhibitors also reduced autophagy. Nitric oxide or H(2)O(2) induced autophagy in cardiomyocytes, whereas N-acetyl-cysteine, a potent antioxidant, suppressed autophagy. LPS resulted in increased reactive oxygen species (ROS) production and decreased total glutathione. To test the hypothesis that autophagy might serve as a damage control mechanism to limit further ROS production, we induced autophagy with rapamycin before LPS exposure. The activation of autophagy by rapamycin suppressed LPS-mediated ROS production and protected cells against LPS toxicity. These findings support the notion that autophagy is a cytoprotective response to LPS-induced cardiomyocyte injury; additional studies are needed to determine the therapeutic implications.
细菌内毒素脂多糖(LPS)是脓毒症休克特征性多器官功能障碍的病因,也是患者内毒素血症常见特征——心肌抑制的病因。在这种情况下,心肌功能障碍似乎部分归因于促炎细胞因子的产生。一系列证据还表明,LPS刺激心肌细胞自噬。然而,导致自噬的信号转导途径及其在心脏中的作用尚未完全明确。在本研究中,我们希望确定LPS对自噬的影响以及自噬反应的生理意义。在给予细菌LPS或TNF-α后,对HL-1心肌细胞、新生大鼠心肌细胞和转基因小鼠心脏的自噬进行形态学和生化监测。我们观察到,暴露于LPS或由LPS诱导的TNF-α后自噬增加。AG126对TNF-α产生的抑制显著减少了细胞培养和体内自噬体的积累。药理学抑制剂对p38丝裂原活化蛋白激酶或一氧化氮合酶的抑制也减少了自噬。一氧化氮或H₂O₂诱导心肌细胞自噬,而强效抗氧化剂N-乙酰半胱氨酸抑制自噬。LPS导致活性氧(ROS)产生增加和总谷胱甘肽减少。为了验证自噬可能作为一种损伤控制机制来限制进一步ROS产生的假设,我们在LPS暴露前用雷帕霉素诱导自噬。雷帕霉素激活自噬抑制了LPS介导的ROS产生,并保护细胞免受LPS毒性。这些发现支持自噬是对LPS诱导的心肌细胞损伤的一种细胞保护反应这一观点;需要进一步研究来确定其治疗意义。